Expression of recombinant classical swine fever virus E2 glycoprotein instable High Five cells

Abstract

Classical swine fever is one of the most important viral diseases of domestic pigs and wild boar. It is a notifiable disease to the World Organization for Animal Health. In the present study, codon optimized CSFV E2 gene was cloned into a laboratory-modified insect cell expression vector for producing the recombinant protein in secretory form. Endotoxin-free recombinant plasmid prepared from the positive clone was transfected into the High Five insect cell line and then selected against zeocin antibiotic for the development of a stable cell line. The stable High Five cell line successfully secreted E2 recombinant protein with a molecular weight of ~65 kDa as revealed by SDS-PAGE and Western blot. The recombinant E2 protein showed efficient reactivity with known CSFV anti-serum and monoclonal antibodies (MAbs against E2) in dot blot assay indicating its potential as a diagnostic antigen.

Keywords

• Classical swine fever virus (CSFV), Dot blot, Insect cell expression, Western blot